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A single-tube six-colour flow cytometry screening assay for the detection of minimal residual disease in myeloma

Leukemia volume 21, pages 2046–2049 (2007)Cite this article

Detection of minimal residual disease (MRD) in myeloma is highly predictive of early relapse following treatment. A number of different approaches to myeloma MRD detection are available; these vary widely in sensitivity and cost. Although several studies suggest that detection of neoplastic plasma cells above a level of 0.01% is clinically relevant,1, 2, 3, 4, 5, 6 MRD studies in B-cell chronic lymphocytic leukemia (CLL) comparing different approaches have demonstrated that methods such as consensus-primer immunoglobulin heavy-chain (IgH) PCR and assessment of light-chain restriction by flow cytometry can detect low levels of disease but the sensitivity is highly variable. Such approaches are least informative when large numbers of polyclonal B cells are present as is typical after high-dose therapy and stem cell transplantation. In this setting, false-negative results may be generated in cases with up to 5% residual disease. Therefore, approaches that can directly quantify the proportion of CLL cells, using either CLL-specific flow cytometry assays or real-time quantitative allele-specific oligonucleotide (RQ-ASO) IgH-PCR are recommended for MRD detection as the sensitivity is not impaired by the presence of polyclonal B cells.7, 8

In both CLL and myeloma, flow cytometric assessment of MRD may be preferable in practice to RQ-ASO IgH-PCR since a clinically relevant limit of MRD detection can be achieved using a standard antibody panel whereas RQ-ASO IgH-PCR requires the cost and labour-intensive development of patient-specific primers. However, quantitative MRD detection of myeloma plasma cells by flow cytometry is relatively complicated compared to CLL flow cytometric MRD analysis. Myeloma MRD flow cytometry requires at least two markers for plasma cell identification (CD38 and either CD45 or CD138, preferably all three) and some combination of several additional markers to detect phenotypic abnormality including CD19, CD20, CD27, CD28, CD45, CD56 and CD117.4, 5, 6, 9, 10 As most of the myeloma-associated phenotypic plasma cell abnormalities can also be detected at low levels in healthy individuals, assessment of immunoglobulin light-chain restriction (cytoplasmic κ and λ) is also informative.

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Acknowledgements

We thank Medical Research Council (MRC), UK and Leukaemia Research Fund (LRF), UK. RMdT and ACR are members of the EuroFlow consortium.

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  1. HMDS, Leeds Teaching Hospitals NHS Trust, Leeds, UK

    R M de Tute, A S Jack, J A Child, R G Owen & A C Rawstron

  2. Department of Haemato-oncology, Royal Marsden Hospital, Surrey, UK

    G J Morgan

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Correspondence to A C Rawstron.

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de Tute, R., Jack, A., Child, J. et al. A single-tube six-colour flow cytometry screening assay for the detection of minimal residual disease in myeloma. Leukemia 21, 2046–2049 (2007). https://doi.org/10.1038/sj.leu.2404815

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